Identification and characterization of the murine Rag1 promoter.
Rag1 and Rag2 are required for the somatic rearrangement of immunoglobulin genes and T-cell receptor genes and the subsequent development of B and T cells. We describe the pattern of DNase I hypersensitive sites surrounding the Rag1 gene that accompanies mouse B-cell development and show that one of these sites corresponds to the murine Rag1 promoter. Transcription initiates over a 30 bp region, with approximately 70% of the transcripts initiating within a 5 bp region. The promoter contains neither a consensus TATA box nor an initiator, but does contain an AT rich sequence that could serve as a non-consensus TATA box. The Rag1 promoter directs only negligible levels of expression in transient transfection assays, but when combined with a heterologous enhancer, it is capable of driving significant levels of expression in pre-B cells, pre-T cells, and mature B cells. Methylation interference and mutation analysis reveal that the Rag1 promoter contains binding sites for E-box binding proteins, NF-Y proteins, and Ikaros proteins. These findings are discussed with respect to B-cell development and regulation of differential Rag expression by the promoter in pre-B, pre-T, and CNS cells.
Fuller
K
K
Department of Molecular Genetics and Cell Biology, The University of Chicago, IL 60637, USA.
Storb
U
U
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics*; D015322:Gene Rearrangement, B-Lymphocyte; D015329:Gene Rearrangement, T-Lymphocyte; D016654:Genes, RAG-1; D006041:Goats; D018398:Homeodomain Proteins; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C064658:RAG-1 protein